Flow electroporation chamber and method

• US 6,773,669 B1
• Filed: 11/08/2000
• Issued: 08/10/2004
• Est. Priority Date: 03/10/1995
• Status: Expired due to Fees

• Current Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Original Assignee: Maxcyte, Inc. (Entremed, Incorporated)
• Inventors: Holaday, John W., Singh, Vininder, Meserol, Peter H., Doerfler, Doug, Green, Shawn J.
• Primary Examiner(s): NGUYEN, DAVE TRONG
• Application Number:
  US09/707,928
• Time in Patent Office: 1,371 Days
• Field of Search: 435/172.3, 435/173.6, 422/44, 422/33
• US Class Current: 422/44
• CPC Class Codes: A61K 35/18 Erythrocytes haemoglobin A61K38/42
• CPC Class Codes: A61M 1/16 with membranes
• CPC Class Codes: A61M 1/3687 Chemical treatment A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2202/0092 starting product created by centrifuging
• CPC Class Codes: A61M 2202/0429 Red blood cells; Erythrocytes
• CPC Class Codes: A61M 2205/3306 Optical measuring means
• CPC Class Codes: A61M 2205/3368 Temperature
• CPC Class Codes: A61M 2205/3382 Upper level detectors
• CPC Class Codes: A61M 2205/3606 cooled
• CPC Class Codes: A61M 2205/3673 thermo-electric, e.g. Peltier effect, thermocouples, semi-conductors
• CPC Class Codes: A61M 2205/50 with microprocessors or computers
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: F25B 21/02 using Peltier effect; using Nernst-Ettinghausen effect

Abstract

The present invention relates to a method and apparatus for the encapsulation of substances and drugs into cells and platelets. The present invention is also related to the incorporation of thrombus dissolving drugs, such as tissue plasminogen activator and streptokinase into platelets using the apparatus described herein. The treated platelets can then be used to treat patients suffering from a thrombus blocking a blood vessel. The present invention is also related to a preparation of red blood cells that has a stable right shift of the oxygen dissociation curve.

Claims

1. An apparatus for electroporation, comprising:

a fluid flow path;

electrodes disposed along sides of the fluid flow path and configured to subject biological particles moving along the fluid flow path to an electrical field suitable for electroporation; and

a computer-controllable pump in fluid communication with the fluid flow path and configured to establish a flow rate of the fluid flow path in accordance with a sample processing rate.

2. The apparatus of claim 1, wherein the electrical energy is pulsed.

3. The electroporation chamber of claim 1, wherein the electrical energy is a variable flux.

4. The apparatus of claim 1, wherein the electrodes comprise continuous band electrodes.

5. The apparatus of claim 1, further comprising thermoelectric cooling elements in operative relation with the electrodes.

11. The apparatus of claim 1, the computer-controllable pump configured to vary the flow rate of the fluid flow path to match the sample processing rate.

12. The apparatus of claim 1, the sample processing rate corresponding to a processing rate of a centrifuge.

13. The apparatus of claim 1, the sample processing rate corresponding to an interval between pulses of electrical energy delivered to the electrodes.

14. The apparatus of claim 1, where one of the walls defining the fluid flow path is elastically deformable and at least another one of the walls defining the fluid flow path is substantially rigid.

6. An apparatus for electroporation comprising:

walls defining a fluid flow path;

electrodes disposed along sides of the fluid flow path and configured to subject biological particles moving along the fluid flow path to an electrical field suitable for electroporation; and

means for breaking the electrodes prior to the apparatus being removed from the support member whereby the apparatus cannot be re-used.

7. The apparatus of claim 6, wherein:

the electrodes are wrapped around at least a portion of a spindle; and

the means for breaking comprise spindles configured to rotate so as to stretch the electrodes beyond their tensile limits to break the electrodes and render them electrically inoperative.

8. An apparatus for electroporation comprising:

walls defining a fluid flow path;

electrodes disposed along sides of the fluid flow path and configured to subject biological particles moving along the fluid flow path to an electrical field suitable for electroporation;

a pump for moving the biological particles along the fluid flow path; and

a computer responsive to the rate at which the pump moves the biological particles along the fluid flow path and to the interval between pulses of electrical energy.

9. The apparatus of claim 8, wherein the computer regulates the rate at which the pump moves the biological particles along the fluid flow path.

10. The apparatus of claim 8, wherein the computer regulates the interval between pulses of electrical energy.

15. An apparatus for electroporation, comprising:

a fluid flow path;

electrodes coupled to the fluid flow path and configured to subject biological particles moving along the fluid flow path to an electrical field suitable for electroporation;

a pump configured to establish a flow rate of the fluid flow path; and

a computer configured to;

a. control charging of the electrodes; and

b. establish a flow rate of the fluid flow path in accordance with a sample processing rate.

16. The apparatus of claim 15, the sample processing rate corresponding to a processing rate of a centrifuge.

17. The apparatus of claim 15, the sample processing rate corresponding to an interval between pulses of electrical energy delivered to the electrodes.

18. The apparatus of claim 15, the computer configured to control the thermoelectric cooling elements individually to cool one location more or less than another location.

19. The apparatus of claim 15, further comprising:

valves configured to control fluid flow;

thermoelectric cooling elements; and

wherein the computer is further configured to;

c. control operation of the valves;

or

d. control the thermoelectric cooling elements.